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Some patients develop persistent eye pain after refractive surgery, but factors that cause or sustain pain are unknown. We tested whether tear proteins of patients with pain 3 months after surgery differ from those of patients without pain. Patients undergoing refractive surgery (laser in situ keratomileusis or photorefractive keratectomy ) were recruited from 2 clinics, and tears were collected 3 months after surgery. Participants rated their eye pain using a numerical rating scale (NRS, 0–10; no pain–worst pain) at baseline, 1 day, and 3 months after surgery. Using tandem mass tag proteomic analysis, we examined tears from patients with pain [NRS ≥ 3 at 3 months (n= 16)] and patients with no pain [NRS ≤ 1 at 3 months (n= 32)] after surgery. A subset of proteins (83 of 2748 detected, 3.0%) were associated with pain 3 months after surgery. High-dimensional statistical models showed that the magnitude of differential expression was not the only important factor in classifying tear samples from pain patients. Models utilizing 3 or 4 proteins had better classification performance than single proteins and represented differences in both directions (higher or lower in pain). Thus, patterns of protein differences may serve as biomarkers of postsurgical eye pain as well as potential therapeutic targets.

IMPORTANCE: Optimal strategies for population-based outreach for breast cancer screening remain unknown. OBJECTIVE: To evaluate the effect on breast cancer screening of an opt-out automatic mammography referral strategy compared with an opt-in automated telephone message strategy. DESIGN, SETTING, AND PARTICIPANTS: This pragmatic randomized clinical trial was conducted from April 2022 to January 2023 at a single Veterans Affairs (VA) medical center. Participants were female veterans aged 45 to 75 years who were eligible for breast cancer screening and enrolled in VA primary care. INTERVENTION: Veterans were randomized 1:1 to receive either an automatic mammography referral (opt-out arm) or an automated telephone call with an option for mammography referral (opt-in arm). MAIN OUTCOMES AND MEASURES: The primary outcome was completed mammography 100 days after outreach. Secondary outcomes were scheduled or completed mammography by 100 days after outreach and referrals canceled if mammography was not scheduled within 90 days. Both intention-to-treat analyses and a restricted analysis were conducted. The restricted analysis excluded veterans who were unable to be reached by telephone (eg, a nonworking number) or who were found to be ineligible after randomization (eg, medical record documentation of recent mammography). RESULTS: Of 883 veterans due for mammography (mean [SD] age, 59.13 [8.24] years; 656 [74.3%] had received prior mammography), 442 were randomized to the opt-in group and 441 to the opt-out group. In the intention-to-treat analysis, there was no significant difference in the primary outcome of completed mammography at 100 days between the opt-out and opt-in groups (67 [15.2%] vs 66 [14.9%]; P = .90) or the secondary outcome of completed or scheduled mammography (106 [24.0%] vs 84 [19.0%]; P = .07). A higher number of referrals were canceled in the opt-out group compared with the opt-in group (104 [23.6%] vs 24 [5.4%]; P < .001). The restricted analysis demonstrated similar results except more veterans completed or scheduled mammography within 100 days in the opt-out group compared with the opt-in group (102 of 388 [26.3%] vs 80 of 415 [19.3%]; P = .02). CONCLUSIONS AND RELEVANCE: In this randomized clinical trial, an opt-out population-based breast cancer screening outreach approach compared with an opt-in approach did not result in a significant difference in mammography completion but did lead to substantially more canceled mammography referrals, increasing staff burden. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT05313737

Genome-wide association studies indicate that allele variants in MIR137, the host gene of microRNA137 (miR137), confer an increased risk of schizophrenia (SCZ). Aberrant expression of miR137 and its targets, many of which regulate synaptic functioning, are also associated with an increased risk of SCZ. Thus, miR137 represents an attractive target aimed at correcting the molecular basis for synaptic dysfunction in individuals with high genetic risk for SCZ. Advancements in nanotechnology utilize lipid nanoparticles (LNPs) to transport and deliver therapeutic RNA. However, there remains a gap in using LNPs to regulate gene and protein expression in the brain. To study the delivery of nucleic acids by LNPs to the brain, we found that LNPs released miR137 cargo and inhibited target transcripts of interest in neuroblastoma cells. Biodistribution of LNPs loaded with firefly luciferase mRNA remained localized to the mouse prefrontal cortex (PFC) injection site without circulating to off-target organs. LNPs encapsulating Cre mRNA preferentially co-expressed in neuronal over microglial or astrocytic cells. Using quantitative proteomics, we found miR137 modulated glutamatergic synaptic protein networks that are commonly dysregulated in SCZ. These studies support engineering the next generation of brain-specific LNPs to deliver RNA therapeutics and improve symptoms of central nervous system disorders.

Understanding which factors predict primary care provider (PCP) turnover can help organizations prepare for PCP shortages. We conducted a retrospective cohort study of Veteran Health Administration PCPs between 2012 and 2016. We analyzed whether 7 domains of the patient centered medical home (PCMH) implementation—including access, care coordination, comprehensiveness, self-management support, communication, shared decision-making, and team-based care—were associated with PCP turnover. We found that 2 domains of PCMH (access and self-management) were associated with lower turnover, which may reflect that practice cultures that support these characteristics may lower PCP turnover.

•OxLDL enhances GPVI-driven platelet procoagulant activity ex vivo.•P2YR antagonists and tyrosine kinase inhibitors may reduce oxLDL-enhanced platelet procoagulant activity more so than aspirin.

Polymer nanocomposites (PNCs) have showed the significant applications in the fields of packing, energy, safety, defense, sensors, EMI shielding, optoelectronics etc. Tungsten trioxide nanoparticles (NPs) were prepared by propellant chemistry technique using Azadirachta indicaextracts as a fuel/surfactant. Conducting polyaniline (PANI) was prepared using aniline monomer and suitable polymerizing agents. Ex-situ nanocomposites were prepared by mixing the different weight percentage of NPs into PANI matrix. For the better dispersion of NPs in to the matrix, solvents like Dodesyl benzene sulphonic acid (DBSA) and Camphor sulphonic acid (CSA) were used. Crystal structure, morphological features and their dielectric response at room temperature were reported. Electromagnetic Shielding (EMI) studies were compared between the PANI-WO3composites prepared by CSA and DBSA solvents. Composites showed worthy shielding response of 80 dB with CSA and 38 dB with DBSA. Here, the protonation of the solvent and the dispersion of NPs with the polymer matrix matters for the shielding efficiency. Hence, prepared materials are suitable for packing of electronic devices which provides the better EMI shielding and also suppress the heat through effective absorption of EM waves of X-Band.

Ex vivoassays of platelet function critically inform mechanistic and clinical hematology studies, where effects of divergent blood processing methods on platelet composition are apparent, but unspecified.

Ex vivoassays of platelet function critically inform mechanistic and clinical hematology studies, where effects of divergent blood processing methods on platelet composition are apparent, but unspecified. Here, we evaluate how different blood anticoagulation options and processing times affect platelet function and protein content ex vivo. Parallel blood samples were collected from healthy human donors into sodium citrate, acid citrate dextrose, EDTA or heparin, and processed over an extended time course for functional and biochemical experiments, including platelet proteome quantification with multiplexed tandem mass tag (TMT) labeling and triple quadrupole mass spectrometry (MS). Each anticoagulant had time‐dependent effects on platelet function in whole blood. For instance, heparin enhanced platelet agonist reactivity, platelet‐monocyte aggregate formation and platelet extracellular vesicle release, while EDTA increased platelet α‐granule secretion. Following platelet isolation, TMT‐MS quantified 3357 proteins amongst all prepared platelet samples. Altogether, >400 proteins were differentially abundant in platelets isolated from blood processed at 24 h versus 1 h post‐phlebotomy, including proteins pertinent to membrane trafficking and exocytosis. Anticoagulant‐specific effects on platelet proteomes included increased complement system and decreased α‐granule proteins in platelets from EDTA‐anticoagulated blood. Platelets prepared from heparinized blood had higher levels of histone and neutrophil‐associated proteins in a manner related to neutrophil extracellular trap (NET) formation and platelet:NET interactions in whole blood ex vivo. Our results demonstrate that different anticoagulants routinely used for blood collection have varying effects on platelets ex vivo, where methodology‐associated alterations in platelet proteome may influence mechanistic, translational and biomarker studies.

To determine if the secretions collected from a conditionally reprogrammed primary endocervical cell culture are suitable surrogates for mucus studies.

Shopper buying behaviour is essential for the retailers to segment the shoppers in accordance to their disruptive attitude and perception for better innovative strategies which may lead to higher profits. The major purpose of this study to categorize the shoppers into distinct groups based on their risk-based perception for the organized retail outlets in Bangladesh. Seven hundred eighty-five respondents were responding on 21 variables related to store which influence their buying behaviour. In the present study, the shoppers were classified into three segments such as value seekers and disruptive to please shoppers, quality and style-driven shoppers, sensory-driven, and not interested shoppers by using innovative k-means cluster analysis. The results of the study help to retailers in understanding the various disruptive segments of shoppers in relation to their importance for store attributes affected by their demographic characteristics and guide the retailers to take necessary actions regard redesign of retail mix to provide innovative value to the shoppers.

Platelets engage cues of pending vascular injury through coordinated adhesion, secretion, and aggregation responses. These rapid, progressive changes in platelet form and function are orchestrated downstream of specific receptors on the platelet surface and through intracellular signaling mechanisms that remain systematically undefined. This study brings together cell physiological and phosphoproteomics methods to profile signaling mechanisms downstream of the immunotyrosine activation motif (ITAM) platelet collagen receptor GPVI. Peptide tandem mass tag (TMT) labeling, sample multiplexing, synchronous precursor selection (SPS), and triple stage tandem mass spectrometry (MS3) detected >3000 significant (false discovery rate < 0.05) phosphorylation events on >1300 proteins over conditions initiating and progressing GPVI-mediated platelet activation. With literature-guided causal inference tools, >300 site-specific signaling relations were mapped from phosphoproteomics data among key and emerging GPVI effectors (ie, FcRγ, Syk, PLCγ2, PKCδ, DAPP1). Through signaling validation studies and functional screening, other less-characterized targets were also considered within the context of GPVI/ITAM pathways, including Ras/MAPK axis proteins (ie, KSR1, SOS1, STAT1, Hsp27). Highly regulated GPVI/ITAM targets out of context of curated knowledge were also illuminated, including a system of >40 Rab GTPases and associated regulatory proteins, where GPVI-mediated Rab7 S72 phosphorylation and endolysosomal maturation were blocked by TAK1 inhibition. In addition to serving as a model for generating and testing hypotheses from omics datasets, this study puts forth a means to identify hemostatic effectors, biomarkers, and therapeutic targets relevant to thrombosis, vascular inflammation, and other platelet-associated disease states.

Platelets engage cues of pending vascular injury through coordinated adhesion, secretion, and aggregation responses. These rapid, progressive changes in platelet form and function are orchestrated downstream of specific receptors on the platelet surface and through intracellular signaling mechanisms that remain systematically undefined. This study brings together cell physiological and phosphoproteomics methods to profile signaling mechanisms downstream of the immunotyrosine activation motif (ITAM) platelet collagen receptor GPVI. Peptide tandem mass tag (TMT) labeling, sample multiplexing, synchronous precursor selection (SPS), and triple stage tandem mass spectrometry (MS3) detected >3000 significant (false discovery rate < 0.05) phosphorylation events on >1300 proteins over conditions initiating and progressing GPVI-mediated platelet activation. With literature-guided causal inference tools, >300 site-specific signaling relations were mapped from phosphoproteomics data among key and emerging GPVI effectors (ie, FcRγ, Syk, PLCγ2, PKCδ, DAPP1). Through signaling validation studies and functional screening, other less-characterized targets were also considered within the context of GPVI/ITAM pathways, including Ras/MAPK axis proteins (ie, KSR1, SOS1, STAT1, Hsp27). Highly regulated GPVI/ITAM targets out of context of curated knowledge were also illuminated, including a system of >40 Rab GTPases and associated regulatory proteins, where GPVI-mediated Rab7 S72phosphorylation and endolysosomal maturation were blocked by TAK1 inhibition. In addition to serving as a model for generating and testing hypotheses from omics datasets, this study puts forth a means to identify hemostatic effectors, biomarkers, and therapeutic targets relevant to thrombosis, vascular inflammation, and other platelet-associated disease states.

Metal granshot is a process that is used to convert metal lumps or scraps into metal grains of uniform sizes and shapes. The concept of metal granulation is based on the heat difference between melting and pouring medium. The commercial uses of metal granules are jewelry making and spray coating of costly metals such as silver, gold, and platinum. The basic aim of the silver granshot process is to get uniform size, shape, and maximum yield of silver metal granules. During processing, the metal gets superheated within vacuum up to viscosity more than 2.2 cP flow out of the minimum diameter orifice and allow to drop in the coolant with high pressurized argon gas. The present study focuses on the granulation of silver and tries to achieve the maximum yield of metal. The shape and size of the granules produced were observed using scanning electron microscopy and the results show that the average diameters of granules are between 1.2 mm and 3.8 mm. Yield is also considered as an important factor for granulation of valuable metals like silver, and after experimentation successfully achieved the 99.9 percent yield. The granulation process of silver will help to ease the trading of metal in the jewelry industry.

Supplemental Digital Content is available in the text.

Aging is an independent risk factor for the development of cardiovascular, thrombotic, and other chronic diseases. However, mechanisms of platelet hyperactivation in aging remain poorly understood.

Supplemental Digital Content is available in the text.

The lack of high-throughput methods to analyze the adipose tissue protein composition limits our understanding of the protein networks responsible for age and diet related metabolic response. We have developed an approach using multiple-dimension liquid chromatography tandem mass spectrometry and extended multiplexing (24 biological samples) with tandem mass tags (TMT) labeling to analyze proteomes of epididymal adipose tissues isolated from mice fed either low or high fat diet for a short or a long-term, and from mice that aged on low versushigh fat diets. The peripheral metabolic health (as measured by body weight, adiposity, plasma fasting glucose, insulin, triglycerides, total cholesterol levels, and glucose and insulin tolerance tests) deteriorated with diet and advancing age, with long-term high fat diet exposure being the worst. In response to short-term high fat diet, 43 proteins representing lipid metabolism (e.g.AACS, ACOX1, ACLY) and red-ox pathways (e.g.CPD2, CYP2E, SOD3) were significantly altered (FDR < 10%). Long-term high fat diet significantly altered 55 proteins associated with immune response (e.g.IGTB2, IFIT3, LGALS1) and rennin angiotensin system (e.g.ENPEP, CMA1, CPA3, ANPEP). Age-related changes on low fat diet significantly altered only 18 proteins representing mainly urea cycle (e.g.OTC, ARG1, CPS1), and amino acid biosynthesis (e.g.GMT, AKR1C6). Surprisingly, high fat diet driven age-related changes culminated with alterations in 155 proteins involving primarily the urea cycle (e.g.ARG1, CPS1), immune response/complement activation (e.g.C3, C4b, C8, C9, CFB, CFH, FGA), extracellular remodeling (e.g.EFEMP1, FBN1, FBN2, LTBP4, FERMT2, ECM1, EMILIN2, ITIH3) and apoptosis (e.g.YAP1, HIP1, NDRG1, PRKCD, MUL1) pathways. Using our adipose tissue tailored approach we have identified both age-related and high fat diet specific proteomic signatures highlighting a pronounced involvement of arginine metabolism in response to advancing age, and branched chain amino acid metabolism in early response to high fat feeding. Data are available via ProteomeXchange with identifier PXD005953.

Primary care intensive management programs utilize interdisciplinary care teams to comprehensively meet the complex care needs of patients at high risk for hospitalization. The mixed evidence on the effectiveness of these programs focuses on average treatment effects that may mask heterogeneous treatment effects (HTEs) among subgroups of patients. We test for HTEs by patients’ demographic, economic, and social characteristics.

IMPORTANCE: Primary care provider (PCP) turnover is common and can disrupt patient continuity of care. Little is known about the effect of PCP turnover on patient care experience and quality of care. OBJECTIVE: To measure the effect of PCP turnover on patient experiences of care and ambulatory care quality. DESIGN, SETTING, AND PARTICIPANTS: Observational, retrospective cohort study of a nationwide sample of primary care patients in the Veterans Health Administration (VHA). We included all patients enrolled in primary care at the VHA between 2010 and 2012 included in 1 of 2 national data sets used to measure our outcome variables: 326 374 patients in the Survey of Healthcare Experiences of Patients (SHEP; used to measure patient experience of care) associated with 8441 PCPs and 184 501 patients in the External Peer Review Program (EPRP; used to measure ambulatory care quality) associated with 6973 PCPs. EXPOSURES: Whether a patient experienced PCP turnover, defined as a patient whose provider (physician, nurse practitioner, or physician assistant) had left the VHA (ie, had no patient encounters for 12 months). MAIN OUTCOMES AND MEASURES: Five patient care experience measures (from SHEP) and 11 measures of quality of ambulatory care (from EPRP). RESULTS: Nine percent of patients experienced a PCP turnover in our study sample. Primary care provider turnover was associated with a worse rating in each domain of patient care experience. Turnover was associated with a reduced likelihood of having a positive rating of their personal physician of 68.2% vs 74.6% (adjusted percentage point difference, −5.3; 95% CI, −6.0 to −4.7) and a reduced likelihood of getting care quickly of 36.5% vs 38.5% (adjusted percentage point difference, −1.1; 95% CI, −2.1 to −0.1). In contrast, PCP turnover was not associated with lower quality of ambulatory care except for a lower likelihood of controlling blood pressure of 78.7% vs 80.4% (adjusted percentage point difference, −1.44; 95% CI, −2.2 to −0.7). In 9 measures of ambulatory care quality, the difference between patients who experienced no PCP turnover and those who had a PCP turnover was less than 1 percentage point. These effects were moderated by the patients’ continuity with their PCP prior to turnover, with a larger detrimental effect of PCP turnover among those with higher continuity prior to the turnover. CONCLUSIONS AND RELEVANCE: Primary care provider turnover was associated with worse patient experiences of care but did not have a major effect on ambulatory care quality.

This study used a proteomics approach to characterize and quantify multipotent adult progenitor cell (MAPC) secretome components in vitro in the presence of inflammatory triggers or a tolerogenic CD74 ligand. Findings provide a background for experiments assessing MAPC response to molecular and cellular triggers and events, for eventual programming of MAPCs to be used in a wide range of clinical applications. Multipotent adult progenitor cells (MAPCs) are adult adherent stromal stem cells currently being assessed in acute graft versus host disease clinical trials with demonstrated immunomodulatory capabilities and the potential to ameliorate detrimental autoimmune and inflammation‐related processes. Our previous studies documented that MAPCs secrete factors that play a role in regulating T‐cell activity. Here we expand our studies using a proteomics approach to characterize and quantify MAPC secretome components secreted over 72 hours in vitro under steady‐state conditions and in the presence of the inflammatory triggers interferon‐γ and lipopolysaccharide, or a tolerogenic CD74 ligand, RTL1000. MAPCs differentially responded to each of the tested stimuli, secreting molecules that regulate the biological activity of the extracellular matrix (ECM), including proteins that make up the ECM itself, proteins that regulate its construction/deconstruction, and proteins that serve to attach and detach growth factors from ECM components for redistribution upon appropriate stimulation. MAPCs secreted a wide array of proteases, some detectable in their zymogen forms. MAPCs also secreted protease inhibitors that would regulate protease activity. MAPCs secreted chemokines and cytokines that could provide molecular guidance cues to various cell types, including neutrophils, macrophages, and T cells. In addition, MAPCs secreted factors involved in maintenance of a homeostatic environment, regulating such diverse programs as innate immunity, angiogenesis/angiostasis, targeted delivery of growth factors, and the matrix‐metalloprotease cascade.

Background: HIV treatment programs in Africa typically approach all enrolling patients uniformly. Growing numbers of patients are antiretroviral experienced. Defining patients on the basis of antiretroviral experience may inform enrollment practices, particularly if medical outcomes differ. Methods: Baseline and follow-up measures (CD4, weight change, and survival) were compared in a retrospective analysis between antiretroviral-naive (ARV-N) and antiretroviral experience (ARV-E) patients enrolled at the Coptic Hope Center for Infectious Diseases in Nairobi, Kenya and followed between January 2004 and August 2006. Results: 1,307 ARV-N and 962 ARV-E patients receiving highly active antiretroviral therapy (HAART) were followed for median of 9 months (interquartile range: 4-16 months). Compared to ARV-N, ARV-E had substantially higher CD4 count (median cells/mm3, 193 versus 95, P < 0.001) and weight (median kg, 62 versus 57, P < 0.001) at baseline, and lower rates of change in CD4 (-9.2 cells/mm3/month; 95 CI, -11.4 - -7.0) and weight (-0.24 kg/month; 95 CI, -0.35 - - 0.14) over 12 months. Mortality was significantly higher in ARV-N than ARV-E (P 0.001). Conclusions: ARV-E patients form a growing group that differs significantly from ARV-N patients and requires a distinct approach from ARV-N clients. Systematic approaches to streamline care of ARV-E patients may allow focused attention on early ARV-N clients whose mortality risks are substantially higher.

Objectives:A cohort of patients with small primary (T1–T2) squamous cell carcinoma of the oropharynx and advanced cervical nodal metastasis were treated with initial neck dissection(s) followed by definitive radiation therapy with or without chemotherapy. Our rationale for this algorithm and our results are examined.

Using zebrafish, Daniorerio, initial pioneering work in the 1960s revealed carcinogen responsiveness of fish, yet very few subsequent tumorigenesis investigations have utilized this species. We exposed embryos (60 hours postfertilization) and fry (3 week posthatch) to 7,12-dimethylbenz[a]anthracene (DMBA) by immersion in aqueous solutions for 24 hours, at concentrations of 0-1 or 0-5 ppm (mg/L), respectively. Juvenile zebrafish 2 months posthatch were fed a diet containing 0-1,000 ppm DMBA for 4 months. Fish were sampled for histologic evaluation at 7-12 months after the onset of carcinogen treatment. Fry were most responsive to DMBA and showed the widest diversity of target tissues and histologic types of neoplasia, having several types of epithelial, mesenchymal, and neural neoplasia. The principal target tissues for carcinogenic response were liver following embryo or fry exposure, with gill and blood vessel the second and third most responsive tissues in fry. Intestine was the primary target and gill a secondary target in fish that received dietary DMBA as juveniles. These studies indicate that young zebrafish are most responsive to DMBA, showing a greater diversity of neoplasm types than rainbow trout. Thus, zebrafish are a valuable model system in which to study mechanistic aspects of the carcinogenesis process.

We exposed embryos (83 hours postfertilizaton) and fry (3 weeks posthatch) to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) by immersion in aqueous solutions of 0-10 ppm for 1 hour (embryo) or 0-2 ppm for 24 hours (fry). Zebrafish embryos were microinjected with MNNG at levels of 0 or 96 ng/egg. Diets containing 0-2,000 ppm MNNG were fed to juvenile zebrafish for 3 months beginning at 2 months posthatch. Fish were sampled for histopathologic study at 6-12 months after initiation of carcinogen exposure. Embryos and fry were both quite responsive to MNNG; however, juvenile zebrafish were remarkably refractory to MNNG-induced neoplasia. Principal target organs in zebrafish treated as embryos with MNNG were liver and testis, with hepatocellular adenoma the most prevalent hepatic neoplasm. A variety of mesenchymal neoplasms occurred in zebrafish following embryo exposure to MNNG, including chondroma, hemangioma, hemangiosarcoma, leiomyosarcoma, and rhabdomyosarcoma. Testis and blood vessels were primary target organs for MNNG following fry exposure, with seminoma, hemangioma, hemangiosarcoma, and various other epithelial and mesenchymal neoplasms occurring. The zebrafish is a responsive, cost-effective lower vertebrate model system in which to study mechanisms of carcinogenesis.

REDDY, A. S., M. K. REEDY, A. V. SEABER, and W. E. GARRETT, JR. Restriction of the injury response following an acute muscle strain. Med. Sci. Sports Exerc.Vol. 25, No. 3, pp. 321–327, 1993. Indirect skeletal muscle injuries have been found to occur exclusively near the myotendinous junction (MTJ). Although muscle cells are syncytial and extend far into the muscle belly, the response to this injury has been shown to be limited to a focal area near the MTJ. This study examined single muscle fibers near their site of rupture in order to document structural changes that may help to explain this limited injury response. A partial, nondisruptive strain injury was created in a New Zealand white rabbit extensor digitorum longus muscle. The muscles were left in vivofor 60 min or 6 h before harvesting. The specimens were divided into four groups of single fibers: ruptured fibers (60 min) attached to muscle belly (group I); ruptured fibers (60 min) attached to tendon (group II); ruptured fibers (6 h) attached to muscle belly (group III); and normal, unstretched fibers (60 min) at the MTJ (group IV) (N= 10 for each group). Sarcomeres closest to the site of fiber rupture in the three injured groups (I, II, and III) were hypercontracted, with lengths well below the physiologic range (I:0.86 ± 0.08 μm, II: 0.96 ± 0.09 μm, III: 0.96 ± 0.21 μm). There was a progressive increase in sarcomere length, which became normal by 300–500 μm away from the site of rupture (I: 2.17 ± 0.48 μm, II: 2.69 ± 0.42 μm, III: 2.08 ± 0.48 μm). The 6-h fibers in group III showed evidence of necrosis before the transition to normal sarcomere spacing occurred. The presence of severe sarcomere hypercontraction in the 60 min muscle fibers and eventual necrosis in the 6-h fibers suggests a progressive response to cell rupture, consistent with well-known effects of calcium entry. However, in all three injured groups, sarcomere length abruptly normalized within 300–500 μm away from the site of rupture. This suggests that following an acute muscle strain an intracellular barrier effectively restricts the injury response to less than 500 μm away from the initial site of rupture.

The zebrafish (Danio rerio) is assuming prominence in developmental genetics research. By comparison, little is known of tumorigenesis and nothing is known of carcinogen metabolism in this species. This study evaluated the ability of zebrafish to metabolize a well-characterized human carcinogen, aflatoxin B1(AFB1), to phase I and phase II metabolites and assessed hepatic AFB1–DNA adductionin vivo.Fish ip injected with 50–400 μg [3H]AFB1/kg body wt displayed a linear dose response for hepatic DNA binding at 24 hr. AFB1–DNA adduct levels among treatments showed no statistical difference over the period from 1 to 21 days after injection, suggesting poor adduct repair in this species. DNA binding in female fish was 1.7-fold higher than that in males (p< 0.01). Anin vitroAFB1metabolism assay verified that zebrafish liver extracts oxidize AFB1to the 8,9-epoxide proximate electrophile (Km= 79.0 ± 16.4 μm,Vmax= 11.7 ± 1.4 pmol/min/mg protein at 28°C). The excretion of AFB1and its metabolites was also examined by HPLC. As is typical of other fish studied, major metabolites excreted were aflatoxicol (AFL) and aflatoxicol–glucuronide (AFL-g), followed by unreacted AFB1. AFL appeared as early as 5 min after injection, whereas AFL-g was a significant metabolite after 18 hr. This study shows thatin vivoadministration of AFB1to zebrafish results in moderate adduction of the carcinogen to liver DNA and that zebrafish have the capacity for both phase I and phase II metabolism of AFB1. The approximate fourfold difference between rainbow trout and zebrafish AFB1–DNA covalent binding index appears insufficient to explain the relative resistance of zebrafish to dietary AFB1hepatocarcinogenicity.

By designing and evaluating health system improvements and providing evidence to clinical decision-makers, embedded researchers are a critical part of a Learning Health System (LHS). In this article, we describe the evolution and mission of the Primary Care Analytics Team (PCAT), an integrated research team within the Veterans Health Administration Office of Primary Care. We discuss challenges and strategies for success in working with clinical operations partners and provide recommendations for other Learning Health Systems units embedded in large integrated health care organizations.

Chronic daily headaches (CDH) are primarily understood from a psychophysiological formulation. A broad biopsychosocial understanding, where there is equal importance given to biological, psychological and socio-cultural factors, is underexplored in headache. Socio-cultural factors, such as gender, socio-economic factors can perpetuate and worsen the condition. For an effective and sustainable intervention, these factors need to be considered. The current study aims to explore and develop a biopsychosocial framework for headache disorders. A convergent parallel mixed methods design was used and participants were recruited from a tertiary referral care hospital, Bengaluru, India. Headache Assessment Sheet, GAD 7, HDRS, PSS and B COPE were used in the quantitative phase. The data was analysed using r software. Qualitative phase of the study, in depth interview guide was used and data analysed thematically. Quantitative phase, 38 participants were recruited. The average age, was 38.02 (±10.17), majority of the participants being females 31 (81.58%). The mean duration of illness was 8.63 (±4.73) years. The anxiety scores positively correlated with pain intensity (r = 0.50 at P≤ .001) and the median anxiety scores varied with photophobia (P≤ .03). The anxiety scores correlated with PSS (r = 0.428 at P≤ .007) and HDRS (r = 0.428 at P≤ .007) scores. Gender variations in coping were seen, avoidant coping having higher median scores in women. (P ≤ .08). In qualitative phase of the study, six participants were recruited. Three main themes emerged from the qualitative phase of the study: headache - an illness, headache factors and the impact.Chronic daily headaches are influenced by biological, psychological, environmental and socio-cultural factors. A bio-psychosocial framework will help to understand and develop targeted interventions.

ABSTRACTA loop-mediated isothermal amplification (LAMP) assay for the rapid detection of cytomegalovirus (CMV) was developed and evaluated. The LAMP assay specifically amplified only CMV DNA, and no cross-reactivity with the DNA of herpes simplex virus type 1, varicella-zoster virus, adenovirus, Aspergillus flavus, or Staphylococcus aureuswas observed. The sequences of the LAMP assay-positive CMV products were perfectly (100%) matched with the CMV sequence deposited in the GenBank database. The sensitivity of the LAMP assay was found to be 10 copies/µl of CMV DNA. Vitreous samples from 40 patients with suspected retinitis were subjected to LAMP and real-time PCR for the detection of CMV. Of 40 patients with suspected viral retinitis, 10 tested positive for CMV by the real-time PCR and LAMP assays. A 100% concordance was observed between the results of the two methods. The LAMP assay is a rapid, highly specific, and sensitive method for the diagnosis of retinitis caused by CMV.

The coronavirus disease 2019 (COVID-19) pandemic challenged health care organizations to develop ways to provide patient care with rapidly changing guidelines and scarce resources. Clinical leaders and informatics specialists partnered to rapidly develop an electronic health record (EHR) template for primary care staff to screen Veterans at Veterans Affairs (VA) Puget Sound. The template prompts categorization of patients by stability and suspicion for COVID-19, and provides just-in-time triaging advice for clinic staff. Each category is a discrete data element and this information was used by leadership to track screening and testing volumes. We found that a brief, practical EHR note template can be quickly adopted to inform guideline-based screening, direct patient care, and conserve resources.

The Comprehensive Primary Care (CPC) initiative was an alternative payment model implemented from 2012 to 2016 to strengthen primary care by enhancing core functions, including access to care. The association between interventions to enhance access and patients' perception of access is unknown. We performed a cross-sectional analysis of CPC practice surveys and CAHPS patient survey responses pertaining to access and timeliness in 2016. There were regional differences in both patients' perceptions of access and interventions to enhance access, but no association between interventions and patients’ perceptions. Practices with fewer clinicians and whose patients had fewer chronic conditions had better perceived access.